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Publication Detail
Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells
  • Publication Type:
    Chapter
  • Authors:
    Menegollo M, Tessari I, Bubacco L, Szabadkai G
  • Publisher:
    Springer Nature
  • Publication date:
    01/01/2019
  • Place of publication:
    New York (NY), USA
  • Pagination:
    223, 232
  • Volume:
    1925
  • Chapter number:
    19
  • Series:
    Methods in Molecular Biology
  • Editors:
    Raffaello A,Vecellio Reane D
  • ISBN-13:
    978-1-4939-9017-7
  • Status:
    Published
  • Book title:
    Calcium Signalling: Methods and Protocols
  • Language:
    English
  • Keywords:
    Reversed-phase high-performance liquid chromatography (RP-HPLC), ATP/ADP ratio, Energy charge, Glycolysis, Oxidative phosphorylation, Ca²⁺ signal
Abstract
Cytoplasmic and mitochondrial Ca²⁺ signals couple cellular ATP production to activity-related energy demand. In order to accurately determine the bioenergetic effect of Ca²⁺ signals, cellular energy charge, i.e., the compound ratio of the phosphorylated adenine nucleotides AMP, ADP, and ATP, should be estimated. Reversed-phase high-performance liquid chromatography (RP-HPLC) allows the rapid separation and quantitation of these molecules. Here we describe a protocol applied in our laboratories to quantify ATP, ADP, and AMP nucleotides in cellular extracts.
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